Compositions and methods for remediating chemical warfare agent exposed skin

ABSTRACT

The present invention relates to chemical and biological warfare agent decontaminating compositions and methods for using the same to decontaminate animal skin and wounds thereon exposed to the agents. The compositions may comprise a peracid, a hydroperoxide, and a peroxyacid.

CROSS-REFERENCE TO RELATED APPLICATION

This application claims priority benefit under 35 U.S.C. § 120 as aContinuation In Part of International Patent Application No.PCT/US21/12540 filed Jan. 7, 2021, which in turn claims priority under35 U.S.C. § 120 to U.S. patent application Ser. No. 16/900,816, filedJun. 12, 2020 and U.S. patent application Ser. No. 16/736,546, filedJan. 7, 2020. U.S. patent application Ser. No. 16/900,816 is aContinuation in Part of U.S. patent application Ser. No. 16/736,546,which is a Continuation In Part of International Patent Application No.PCT/US2018/041163, filed Jul. 7, 2018, which claims priority under 35U.S.C. § 119(e) to U.S. Provisional Application Ser. No. 62/530,045filed Jul. 7, 2017. U.S. patent application Ser. No. 16/900,816 alsoclaims priority benefit under 35 U.S.C. § 119(e) to U.S. ProvisionalApplication Ser. No. 62/861,810, filed Jun. 14, 2019 and U.S.Provisional Application Ser. No. 63/004,858, filed Apr. 3, 2020. Thedisclosures of all seven applications are incorporated by referenceherein in their entirety.

FIELD OF THE INVENTION

The present invention relates to compositions that are useful forsurface decontamination after chemical or biological warfare agentexposure and methods for producing and using the same. The compositionsmay contain a peroxyacid the corresponding carboxylic acid, and ahydroperoxide.

BACKGROUND OF THE INVENTION

The skin is a primary route of exposure to chemical agents used asweapons of mass destruction. Because of this threat, the U.S. militaryhas invested considerable resources in developing detectors, protectivegarments, and products to remove and/or decontaminate chemical agentexposure to the skin. The currently fielded skin decontamination (DC)product is a lotion known as Reactive Skin Decontamination Lotion(RSDL), which is a mixture of potassium 2,3-butanedione monoximate(KBDO) and diacetylmonoxime (DAM) in a solvent of polyethylene glycolmonomethyl ether (MPEG) and water. RSDL is FDA approved for use on theskin, near eyes, around wounds and equipment against all OP chemicalagents, sulfur mustard and T-2 toxin.1 Military personnel are issuedthree pouches of RSDL; each pouch contains three packets with a spongepad saturated with RSDL. After a suspected exposure to a chemical agent,RSDL is applied by scrubbing the exposed area(s) vigorously with thesponge and allowing it to remain on the skin for at least 2 minutesbefore removing. 2,3 RSDL can be reapplied and left on the skin for upto twenty-four hours. While RSDL is an effective broad spectrum DCproduct, the user community has complained about its expense and some ofthe physical characteristics of the product. This has renewed interestin identifying a more acceptable broad spectrum personal DC product.

SUMMARY OF THE INVENTION

One aspect of this invention provides an aqueous decontaminatingcomposition containing one or more peroxyacids in an amount ranging fromabout 0.0001% to about 20% by weight; one or more hydroperoxides in anamount ranging from about 0.01% to about 25% by weight; and one or morecarboxylic acids in an amount ranging from about 0.01% to about 30% byweight, wherein at least one of the carboxylic acids is the parentcarboxylic acid of the peroxyacid. In one embodiment, at least one ofthe hydroperoxides is hydrogen peroxide. In another embodiment, theamount of the hydroperoxide ranges from about 5% to about 25% by weight.In a further embodiment, the one or more carboxylic acids are selectedfrom C2-10 carboxylic acids, dicarboxylic acids, tricarboxylic acids,β-keto carboxylic acids, and mixtures thereof. In yet a furtherembodiment, the amount of the carboxylic acid ranges from about 5% toabout 22% by weight.

In one embodiment, the peroxyacid is peracetic acid. In anotherembodiment, the peroxyacid is peracetic acid and the parent carboxylicacid is acetic acid. In a further embodiment, the amount of theperoxyacid ranges from about 0.02% to about 6% by weight. In oneembodiment, the composition further contains a stabilizer selected fromthe group consisting of etidronic acid, disodiumethylenediaminetetraacetic acid, ethylenediaminetetraacetic acid,tetrasodium ethylenediaminetetraacetate, citramalic acid, dipicolinicacid, dipicolinic acid N-oxide or a combination thereof. In anotherembodiment, the composition further containing a magnesium salt. In adifferent embodiment, the magnesium salt is the magnesium salt ofperacetic acid. In a further embodiment, the magnesium salt is magnesiumacetate tetrahydrate.

In another embodiment, the composition is an aqueous solutioncontaining: peracetic acid in an amount ranging from less than 1% toabout 10% by weight; hydrogen peroxide in an amount ranging from about5% to about 20% by weight; and acetic acid in an amount ranging fromabout 5% to about 20% by weight.

In another embodiment the decontaminating composition is an aqueouscomposition containing: peracetic acid in an amount ranging from about0.0001% to about 3.5% by weight; hydrogen peroxide in an amount rangingfrom about 0.01% to about 12% by weight; and acetic acid in an amountranging from about 0.01% to about 12% by weight. In one embodiment, thecomposition is capable of decontaminating at least one of human skin,human wounds, or non-human surfaces that have been exposed to a chemicalbiological warfare agent. In another embodiment, the composition iscapable of decontaminating at least one of V-series nerve agents,G-series nerve agents, A-series nerve agents, and mustard agents.

A different aspect of the invention provides a method of decontaminatingskin, wounds, or surfaces, comprising contacting human skin, a humanwound or a non-human surface that has been exposed to a chemical orbiological warfare agent, with an amount of the decontaminatingcomposition that is sufficient to decontaminate the skin, wound orsurface. In another embodiment, the chemical warfare agent is selectedfrom a V-series nerve agent, G-series nerve agent, A-series nerve agent,mustard agent, or a combination thereof. In a further embodiment, thecomposition is formulated in a form selected from the group consistingof a gel, sol, liquid, lotion, irrigation gel, spray, or a combinationthereof. In yet a further embodiment, the decontamination composition iscontacted with the human skin, human wound or non-human surface by atleast one of: soaking the human skin, human wound or nonhuman surface ina solution of the composition, dispensing a pressurized solution of thecomposition onto the human skin or the surface, or applying a coating orlayer of the composition on the human skin, human wound or nonhumansurface.

In another aspect of the invention, a kit is provided in which thecomposition of the present invention is packaged with an absorbentapplicator for the composition or medical countermeasures for chemicalweapons packaged for administering to an exposed subject. In oneembodiment, the absorbent applicator includes an absorbent sponge,natural or synthetic, woven or non-woven fabric, cloth, paper towel ortowelette. In another embodiment, the applicator is saturated with thecomposition prior to packaging. According to another embodiment, theapplication is a mitt.

In another embodiment, the kit further includes a container for safedisposal of used applicators. According to another embodiment, themedical countermeasures include a nerve agent antidote packaged foradministering to an exposed subject.

BRIEF DESCRIPTION OF THE FIGURES

The accompanying drawings, which are incorporated herein and constitutepart of this specification, illustrate the presently preferredembodiments of the invention, and, together with the general descriptionabove and the detailed description given below, serve to explain thefeatures of the invention. In the drawings:

FIG. 1 is a graph of the VX dose lethality curves in animalsdecontaminated with either RSDL or a composition according to theinvention after cutaneous exposure.

FIG. 2 is a depiction of a pulse sequence of modified gradient ¹H-X1D-HSQC for chemical warfare agents (CWAs).

FIG. 3 is a depiction of an NMR spectrum of (a) chemical warfare agentVX and (b) its decontamination product.

FIG. 4 is a graph showing decontamination (DC) of chemical wafare agentHD by various concentrations of a composition according to the inventionfit to a one-phase decay curve.

FIG. 5 is a graph showing decontamination of chemical wafare agent VX byvarious concentrations of a composition according to the invention fitto a one-phase decay curve.

FIG. 6 a depiction of RSDL and a composition according to the invention(2%) skin decontamination (DC) efficacy following topical application ofagent VX. Decontamination was performed 2 min after agent application.The graph shows the probit-predicted dose-lethality curves based on 24hr mortality. The table shows calculated probit estimates, group sizes,and estimated protective ratios (PR).

FIG. 7 is a depiction of skin decontamination efficacy of variousmaterials using a 3-step decontamination method mimicking mass casualtyprocedures, following topical application of VX. The DC was performed 2min after agent application. The graph shows the probit-predicteddose-lethality curves from 24 hr mortality. The table shows calculatedprobit estimates for LD₅₀, 95% CI, slope, group sizes, and estimatedprotective ratios (PR).

DETAILED DESCRIPTION OF THE INVENTION

While various aspects and features of certain embodiments have beensummarized above, the following detailed description illustrates a fewembodiments in further detail to enable one of skill in the art topractice such embodiments. The described examples are provided forillustrative purposes and are not intended to limit the scope of theinvention.

In the following description, for the purposes of explanation, numerousspecific details are set forth in order to provide a thoroughunderstanding of the described embodiments. It will be apparent to oneskilled in the art, however, that other embodiments of the presentinvention may be practiced without some of these specific details.Several embodiments are described herein, and while various features areascribed to different embodiments, it should be appreciated that thefeatures described with respect to one embodiment may be incorporatedwith other embodiments as well. By the same token, however, no singlefeature or features of any described embodiment should be consideredessential to every embodiment of the invention, as other embodiments ofthe invention may omit such features.

The invention relates to compositions that are useful fordecontamination of surfaces exposed to chemical or biological warfareagents. The invention also relates to methods of producing and using thecompositions. The compositions overcome the problems of the prior art byproviding an aqueous composition containing a peroxyacid, thecorresponding carboxylic acid and a hydroperoxide.

The terms “peroxyacid,” “peracid,” “percarboxylic,” and“peroxy-carboxylic acid,” are used interchangeably herein. In general,peroxyacids are compounds of oxidized form of a base organic acid(generally a carboxylic acid) that exist in equilibrium with an oxidizer(generally hydrogen peroxide) and water. Peroxyacids have the formulaR(CO₃H)_(n), where, for example, R is an alkyl, arylalkyl, cycloalkyl,aromatic, or heterocyclic group, and n is one, two, or three, and namedby prefixing the parent acid with “peroxy-.” The R group can besaturated or unsaturated as well as substituted or unsubstituted.Peroxycarboxylic acids can be made by the direct action of an oxidizingagent on a carboxylic acid, by autoxidation of aldehydes, or from acidchlorides, and hydrides, or carboxylic anhydrides with hydrogen orsodium peroxide.

Peroxyacids useful in the compositions and methods of the presentinvention include peroxyformic, peroxyacetic, peroxypropionic,peroxybutanoic, peroxypentanoic, peroxy-hexanoic, peroxyheptanoic,peroxyoctanoic, peroxynonanoic, peroxydecanoic, peroxyundecanoic,peroxydodecanoic, or the peroxyacids of their branched chain isomers,peroxylactic, peroxy-maleic, peroxyascorbic, peroxyhydroxyacetic,peroxyoxalic, peroxymalonic, peroxysuccinic, peroxyglutaric,peroxyadipic, peroxypimelic and peroxy-suberic acid and mixturesthereof.

In some embodiments, the peroxyacid is either a C1 to C11peroxycarboxylic acid, C2 to C6 peroxycarboxylic acid, C1 to C4peroxycarboxylic acid, or a C5 to C11 peroxycarboxylic acid. In someembodiments, the compositions of the invention utilize a combination ofseveral different peroxycarboxylic acids. For example, in someembodiments, the composition includes one or more C1 to C4peroxycarboxylic acids and one or more C5 to C11 peroxycarboxylic acids.In one embodiment, the peroxyacid is peracetic acid (C2), peroxypropionic acid (C3), peroxybutanoic acid (C4), peroxysuccinic andperoxymalonic acid. It should be noted that both the peroxy-succinic andperoxymalonic acid may come from alpha-keto dicarboxylic acids.Furthermore, because these acids exist in the Krebs cycle they aremetabolically active. In some embodiments, the peroxyacid has the samenumber of carbons as one of the carboxylic acids.

In some embodiments, the peroxyacid is peroxyacetic acid. Peroxyacetic(or peracetic) acid is a peroxycarboxylic acid having the formula:CH₃COOH. Generally, peroxyacetic acid is a liquid having an acrid odorat higher concentrations and is freely soluble in water, alcohol, ether,and sulfuric acid.

In some embodiment, the peroxyacid may be a ready-to-use solution or adilatable solution, which enables easy distribution of the composition.The peroxyacid may be any of the peroxyacids described above. In oneembodiment, the peroxyacid is peroxyacetic acid.

In some embodiments, the composition may include at least one peroxyacidat an equilibrium concentration of from 1 ppm to 20 weight percent,based on the weight of the composition. In some embodiments, theperoxyacid may be present at a concentration of less than 5 weightpercent, or less than 1 weight percent. In some embodiments, the amountof peroxyacid ranges from about 1.25% to about 20%, from about 2% toabout 20%, from about 2% to about 10%, from about 2% to about 6%, fromabout 1% to about 10%, from about 1% to about 6%, from about 0.02% toabout 6%, or from about 1.25 to about 3.5%, all by weight of thecomposition. In some embodiments, the composition may be highly diluted,having an amount of peroxyacid ranging from about 0.0001% to 3.5%,0.0001 to 2.5% or 0.0001 to 1%, all by weight.

In addition to the peroxyacid, the composition contains a hydroperoxide.By “hydroperoxide” is meant a compound containing an O₂H group. Examplesof suitable hydroperoxides include, for example, hydrogen peroxide. Inone embodiment, the composition contains hydrogen peroxide, in additionto at least one of methylhydroperoxide or hydroxymethyl hydroperoxide.In some embodiments, the amount of hydroperoxide ranges from about 0.1%to about 25%, from about, 0.1% to about 12%, 5% to about 25%, from about5% to about 20%, from about 7% to about 20%, from about 7% to about 14%,or from about 7% to about 12%, all by weight. When the hydroperoxide ishydrogen peroxide, hydrogen peroxide quantities disclosed herein areexpressed as neat values, and are not quantities of hydrogen peroxide insolution. It should be noted that water in the aqueous compositionincludes water derived from the aqueous hydrogen peroxide used in thecomposition. Any suitable aqueous hydrogen peroxide may be used in thecomposition, such as for example those containing up to 30% by weight,or up to 50% by weight of hydrogen peroxide.

In some embodiments, the composition may also contain a parentcarboxylic acid corresponding to the peroxy acid. As used herein, theterm “parent carboxylic acid” refers to the corresponding carboxylicacid that the peroxyacid is derived from, or is degraded into under atypical storage or production conditions.

The carboxylic acid can be C₂₋₁₀ fatty acid, dicarboxylic acid,tricarboxylic acid, α-keto carboxylic acid, β-keto carboxylic, or amixture thereof. Non-limiting examples of suitable carboxylic acidsinclude, acetic acid, propionic acid, citric acid, succinic acid,glutaric acid, adipic acid, suberic acid, malonic acid, lactic acid,glycolic acid, oxalic acid, pyruvic acid, citramalic acid, acetoaceticacid, citraconic acid, maleic acid, and a mixture thereof.

In some embodiments, the one or more carboxylic acids, individually ortotally, range from about 0.1% to about 30%, from about 0.1% to about12%, from about 5% to about 30%, from about 7% to about 25%, from about7% to about 22%, from about 5% to about 20%, from about 7% to about 20%,or from about 7% to about 12% all by weight in the composition.

In one embodiment, the composition includes one or more hydroperoxidesin an amount ranging from about 0.01% to about 25% by weight; one ormore carboxylic acids in an amount ranging from about 0.01% to about 30%by weight; and one or more peroxyacids in an amount ranging from about0.0001% to about 20% by weight. In one embodiment, the compositionincludes one or more hydroperoxides in an amount ranging from about 5%to about 25% by weight; one or more carboxylic acids in an amountranging from about 7% to about 30% by weight; and one or moreperoxyacids in an amount ranging from about 1% to about 10% by weight.In another embodiment, the composition contains hydrogen peroxide in anamount ranging from about 7% to about 12% by weight; acetic acid in anamount ranging from about 7% to about 12% by weight; and peracetic acidin an amount ranging from less than 1.25% to about 3.5% by weight.

In some embodiments, the composition may also contain one or moreadditional acids, such as for example, at least one of acid, citricacid, succinic acid, glutaric acid, adipic acid, suberic acid, malonicacid, lactic acid, glycolic acid, oxalic acid, pyruvic acid, tartaricacid, formic acid, cis-epoxysuccinic acid, methyltartaric acid, aceticacid, cis-epoxymethylsuccinic acid, maleic acid, citramalic acid,acetoacetic acid, citraconic acid. In some embodiments, the compositionfurther contains at least one of citramalic acid, acetoacetic acid,citraconic acid, maleic acid and any mixture thereof.

In some embodiments, the composition of any of the embodiments mayinclude other compounds. In some embodiments, the composition may alsocontain a bis(hydroperoxide). Suitable bis(hydroperoxides) include, forexample, 3,3-bis(hydroperoxy)butanoic acid,3-bis(hydroperoxy)butaneperoxoic acid, bis(hydroperoxy)propane, orcombinations thereof. It was particularly unexpected that stablecompositions of peroxyacids and bis(hydroperoxides) could be prepared,since peroxyacids are very strong oxidizing agents even at a pH of 2 to8 because the water soluble peracids are decomposing to form freeradicals.

In some embodiments, the composition may also contain an epoxide.

In some embodiments, the composition may also contain a salt oranhydride of the peroxyacid. Suitable salts include, for example, alithium, sodium, potassium, rubidium, cesium, zinc, magnesium, orcalcium salt, or a mixture thereof. The salt can be a magnesium saltsuch as magnesium hydroxide, magnesium carbonate, magnesium salt ofperacetic acid, and the like. In one embodiment, the salt is a magnesiumsalt of peracetic acid, such as, for example, magnesium acetatetetrahydrate.

In one embodiment, the composition also includes at least one oxidizedacetoacetate compound.

In another embodiment the composition also contains a surfactant. In oneembodiment, the amount of surfactant in the composition ranges fromabout 0.1% to about 5.0% by weight, or about 2.0% to about 3.0% byweight. One example of a suitable surfactant is sodium dodecyl sulfate.In one embodiment, a composition according to the present inventioncontains sodium dodecyl sulfate in an amount ranging from about 0.1% toabout 5.0% by weight, about 2.0% to about 3.0% by weight, or about 2.7%by weight. In another embodiment, a composition according to the presentinvention contains about 100 mM sodium dodecyl sulfate.

The composition may be prepared by any suitable method. The methods ofsome embodiments of the invention include contacting at least onecarboxylic acid, such as acetic acid, or a salt of anhydride thereof,and at least one oxidizing agent, such as hydrogen peroxide. Whendescribing a chemical reaction, the terms “treating,” “contacting,” and“reacting” are used interchangeably herein, and refer to adding two ormore reagents under appropriate conditions to produce the indicatedand/or the desired product. The molar ratio of oxidizing agent tocarboxylic acid typically ranges from about 0.5:1 to about 2:1, oftenabout 2:1 to about 6:1. A molar ratio above 1:1 is preferred. The methodproduces a reaction mixture containing at least one peroxyacid at leastone carboxylic acid and at least one hydroperoxide. Exemplary oxidizingagents that are useful in methods of the invention include, but are notlimited to, hydrogen peroxide, barium peroxide, sodium carbonateperoxide, calcium peroxide, sodium perborate, lithium peroxide,magnesium peroxide strontium peroxide, zinc peroxide, potassiumsuperoxide, and the like.

The reaction is generally conducted in an aqueous solution. Othersolvents, such as an organic solvent can also be used in addition to orin place of the aqueous solution. Because it is inexpensive andcommercially available in an aqueous solution, typically hydrogenperoxide is used as an oxidizing agent.

In some embodiments, additional reagents may be used in the reaction,such as, for example acetic anhydride, maleic acid or anhydride,citraconic acid or anhydride, or a mixture thereof.

The methods of other embodiments of the invention involve preparingadmixtures of the key composition components. In one embodiment, theadmixture contains a hydroperoxide, a peroxyacid, and a parentcarboxylic acid corresponding to the peroxy acid. Suitablehydroperoxides are as described herinabove. In one embodiment, theperoxyacid is peracetic acid, the parent carboxylic acid is acetic acid,and the hydroperoxide is hydrogen peroxide. In another embodiment, theadmixture contains hydrogen peroxide, a peroxyacid, such as peraceticacid, and one or more optional compounds such as tartaric acid, formicacid, cis-epoxysuccinic acid, methyltartaric acid,cis-epoxymethylsuccinic acid, maleic acid, citramalic acid or citraconicacid. In one embodiment, the admixture contains hydrogen peroxide,acetic acid and the peroxyacid thereof, peracetic acid.

The compositions of the invention are stable. The composition may be inany suitable, stable form, for example an aerosol or liquid spray or agel, sol, liquid, lotion or irrigation gel, optionally saturated on anabsorbent applicator fabricated all or in part from an absorbentmaterial such as a sponge, natural or synthetic, woven or non-wovenfabric, cloth, paper towel or towelette. The applicator may be suppliedas part of a kit with a container of the composition in which thecomposition is applied to the applicator immediately prior toapplications, or the applicator may be saturated with the compositionprior to packaging. The kit may also include medical countermeasures forchemical weapons, or both, packaged for administering to an exposedsubject, such as a nerve agent antidote. The applicator may be in theform of a mitt. The kit may further include a container for safedisposal of used applicators.

For the purpose of this invention, a “stable” composition is one thatmaintains sufficient physical properties and active oxygen content longenough to be useful, for a period of about twelve months.

In some embodiments, the composition may be useful for at least one ofhealing of wounds after exposure to chemical and biological warfareagents, decontamination of wounds or skin after exposure to chemical andbiological warfare agents, and decontamination of surfaces that havebeen exposed to chemical and biological warfare agents. Chemical andbiological warfare agents are extremely toxic synthetic chemicals thatcan be dispersed as a gas, liquid or aerosol or as agents adsorbed toparticles to become a powder. These chemical and biological warfareagents have either lethal or incapacitating effects on humans.

Chemical warfare agents are usually classified by their effects intonerve agents, blistering agents, blood agents, choking agents,psychomimetic agents (produce changes in thought, perception and mood)and toxins. Among the thousands of toxic substances that are known, onlysome of them are considered chemical warfare agents based on theircharacteristics, including high toxicity, imperceptibility to senses andrapidity of action after dissemination and persistency. Those toxicchemicals qualifying as chemical warfare agents and are listed asscheduled chemicals in the Chemical Weapons Convention (Convention onthe Prohibition of the Development, Production, Stockpiling and use ofChemical Weapons and Destruction, Technical Secretariat of theOrganization for Prohibition of Chemical Weapons, The Hague, accessiblethrough internet. 2005; http://www.opcw.org).

The compositions disclosed herein may be used for decontamination ofessentially any chemical warfare agents. In some embodiments, thecomposition may be useful for decontaminating at least one of V-seriesnerve agents, G-series nerve agents, A-series nerve agents and mustardagents. Examples of V-series agents include VE, VG, VM, VR, VX andanalogues thereof. Examples of G-series agents include GA (tabun), GB(sarin), GD (soman), GF (cyclosarin). Examples of mustard agents includeH, HD, HN₁, NH₂, HN₃, HL, and HT. Examples of A-series agents includeA-230, A-232, A-234, Novichok-5, Novichok-7 and Substance-33. Use of thecomposition for wound decontamination, or skin decontamination, afterchemical agent exposure is particularly important, as the skin is aprimary route of exposure to chemical agents that may be used as weaponsof mass destruction. Likewise, use of the composition fordecontamination of surfaces that have been exposed to chemical orbiological warfare agents is important because the agents remaining onthose surfaces present a risk to persons who touch the contaminatedsurfaces. The composition may be used to decontaminate any surface, suchas for example, the surface of equipment, protective gear, or any othertype of object. In one embodiment, the composition may further include anerve gas antidote. In one embodiment, the nerve gas antidote may beincluded in a kit including the composition.

The composition may be used to decontaminate surfaces by contacting thecomposition with the surfaces, which may include, for example humanskin, human wounds, or non-human surfaces. The contact may be made byany suitable method, for example, by soaking exposed skin, wound or anonhuman surface in a solution of the composition, dispensing apressurized solution of the composition onto exposed skin, wound ornonhuman surface, or applying a coating or layer of the composition ontoexposed skin, wound or nonhuman surface. The coating or layer may beapplied, for example, by wiping or scrubbing with an absorbent sponge,natural or synthetic woven or non-woven fabric, cloth, paper towel ortowelette saturated with the composition of the present invention. Thecomposition may be applied to both human and non-human animal subjectsto decontaminate exposed skin or wounds.

Additional objects, advantages, and novel features of this inventionwill become apparent to those skilled in the art upon examination of thefollowing examples thereof, which are not intended to be limiting. Inthe Examples, procedures that are constructively reduced to practice aredescribed in the present tense, and procedures that have been carriedout in the laboratory are set forth in the past tense.

For clarity, terms used herein are to be understood as described hereinor as such term would be understood by one of ordinary skill in the artof the invention. Additional explanation of certain terms used herein,are provided below:

Unless otherwise indicated, all numbers expressing quantities ofingredients, dimensions, reaction conditions, and so forth, used in thespecification and claims, are to be understood as being modified in allinstances by the term “about”.

In this application and the claims, the use of the singular includes theplural unless specifically stated otherwise. In addition, use of “or”means “and/or” unless stated otherwise. Moreover, the use of the term“including”, as well as other forms, such as “includes” and “included”,is not limiting. Also, terms such as “element” or “component” encompassboth elements and components comprising one unit and elements andcomponents that comprise more than one unit unless specifically statedotherwise.

“wt %” refers to the weight percent relative to the total weight of thesolution or dispersion.

“Film-forming agent” or “water soluble or water dispersible coatingagent,” which may be used interchangeably herein, refer to agents thatform a film and are employed to provide protective coating to thesurface of interest. These agents are either water soluble or waterdispersible. These agents are described in further detail below.

“Locus” as used herein, comprises part or all of a target surfacesuitable to be coated.

While the invention has been particularly shown and described withreference to a number of embodiments, it would be understood by thoseskilled in the art that changes in the form and details may be made tothe various embodiments disclosed herein without departing from thespirit and scope of the invention and that the various embodimentsdisclosed herein are not intended to act as limitations on the scope ofthe claims. All references cited herein are incorporated in theirentirety by reference.

EXAMPLES

The present invention is described more fully by way of the followingnon-limiting examples. Modifications of the examples will be apparent tothose skilled in the art.

Example 1

This example compared the effectiveness of the composition of theinvention as a decontamination (DC) product after skin exposure to thechemical warfare agent VX, as compared to that of Reactive SkinDecontamination Lotion (RSDL), which is a mixture of potassium2,3-butanedione monoximate (KBDO) and diacetylmonoxime (DAM) in asolvent of polyethylene glycol monomethyl ether (MPEG) and water. Thecomposition contained a concentrated solution of 10.7 wt % peraceticacid, 19.8 wt % hydrogen peroxide and 16.8 wt % acetic acid.

Experimental Methods:

Animals: Male guinea pigs [Hartley, Crl(HA)BR] ranging in weight from340-503 gm at the time of experimentation were obtained from CharlesRiver (Canada). After arrival, the animals were maintained in quarantinefor at least 5 days prior to use in an Association for Assessment andAccreditation of Laboratory Animal Care International (AAALACI)accredited animal care and use facility. On the morning of anexperiment, around 0800 hr, animals were weighed, the fur was carefullyremoved from the left side with electric clippers, and excess loose furwas removed with a vacuum. An exposure site was outlined with anindelible marker at approximately the same location on the left side ofeach animal midway between the spine and the ventral midline. Theanimals remained unanesthetized during the entire experiment. Afterexposure to the VX and decontamination with the composition or RSDL,animals were housed in individual cages without bedding in a fume hoodfor the duration of the experiment (24 hr). Food and water were providedad libitum after exposure and DC.

Materials: Each exposure day a 50 pl aliquot of neat VX was obtainedfrom the Chemical Exclusion Area, United States Army Medical ResearchInstitute of Chemical Defense (USAMRICD; Maryland, USA). RSDL waspurchased in sealed packages from First Line Technology, (Chantilly,Va.). The composition according to the invention, containing peracids,was prepared. A 1:6 dilution in deionized water was prepared accordingto the manufacturer's formula each test day.

VX Exposure: Neat VX was applied in a fume hood to the marked exposuresite of each animal, using either a 5 pl Hamilton syringe for volumesgreater than 1 pl, or a 0.5 pl or 1.0 pl Hamilton digital sutyringe forvolumes less than 1 pl. Animals were hand restrained by a trainedtechnician for exposure.

Decontamination procedure: Two minutes after applying VX to the skin,the exposure site was decontaminated with RSDL or the composition.Animals were hand restrained by a trained technician during the DCprocedure. RSDL was applied with an applicator made by stapling ¼ (25mm×50 mm) of a RSDL sponge pad to a wooden tongue depressor. Applicatorsfor the composition were made by stapling a similar size folded gauzepad to wooden tongue depressors. A fresh applicator of each DC productwas used on each animal. The RSDL applicators were made just before thestart of the experiment and were placed into small plastic bags untiluse. The composition applicators were wetted with 10 ml of the dilutedcomposition solution just before DC. Ten ml was sufficient to saturatethe applicator pad without run-off based on previous experience usingdilute bleach or soap and water. RSDL and the composition DC wereperformed by swiping the applicator across the exposure site 10 timesfrom a head to tail direction. Neither DC product was removed afterapplication.

Experimental Design: VX dose-lethality curves were generated for RSDLand the composition, based on 24 hr responses. After exposure and DC,each animal was monitored continuously until the onset of toxic signs,and then at 2 and 4 hr after DC, and again 24 hr after exposure. Amodified stage-wise adaptive dose design was used to generate the VXdose-lethality curves for each DC product. The first stage utilized theclassic up-down dose design of Dixon to estimate the _(LD50) of VX foreach DC product. Briefly, one animal at a time was challenged with adose of VX for each DC product during Stage1. After the 24 hr responsewas determined, the next animal in each DC product group received ahigher (if alive at 24 hr) or lower (if dead at 24 hr) dose of VX,depending on the response of the previous animal. The up-down procedurecontinued until four response reversals were observed. The 24 hrresponses for each DC product from Stage 1 were analyzed by probitanalysis using SAS NLIN and special purpose probit programs developed byBattelle (Columbus, Ohio) to generate an interim LD₅₀ estimate. The nextstages of the experiment used 3-8 animals per stage and various doses ofVX in each stage for each DC product to improve the LD₅₀ estimate andgenerate 95% confidence intervals (CI) by both the Fieller's and thedelta methods. The VX doses in each stage were selected to improve theLD₅₀ estimate and 95% CI based from all stages. Interim probit analyseswere run after each stage, and the experiment was stopped when the ratioof the upper delta 95% CI minus the lower delta 95% CI divided by 2times the LD₅₀ estimate was <0.4. A total of 15 and 26 animals were usedto generate the RSDL and composition dose-lethality curves,respectively.

Statistical Analysis: A final probit analysis was conducted on allstages from the 24 hr responses for RSDL and the composition. Theslopes, LD_(50s) as well as the LD₁, LD10, LD₁₆, LD₃₀, LD₇₀, LD₈₄, LD₉₀,and LD₉₉ with their respective 95% CI were calculated by both Fieller'sand delta methods. Probit estimates were calculated using both targetand actual doses of VX and were not statistically different; therefore,the target doses were used for all statistical comparisons and in thegraphs and tables. LD₅₀ estimates for RSDL and the composition werecompared using SAS and another specialized probit program, whichdetermined whether the ratio of the LD₅₀s was statistically different atp<0.05. A significant (p<0.05) difference was achieved when the delta95% CI of the LD₅₀ ratio did not include the value of 1. The slopes ofthe dose-lethality curves were compared according to Zar. A protectiveratio (PR) defined as LD₅₀ of VX in animals treated with the DC productdivided by the LD₅₀ of VX in untreated animals was estimated, using ahistoric value of 140 μg/kg in fur-clipped unanesthetized guinea pigs(Clarkson, personal communication) for the denominator in the ratio. ThePR expresses the magnitude of the increase in the LD₅₀ by the DCproduct. Another ratio called an absolute efficacy ratio (AER) was alsocalculated. The AER was defined as the LD₁₀ of VX in animals treatedwith a DC product divided by the dermal LD₉₀ of VX in untreated animals.A LD₉₀ value of 188 μg/kg generated in hair-clipped, unanesthetizedguinea pigs (Clarkson, personal communication) was used for thedenominator for the AER. The AER expresses the magnitude of the increasein the LD₁₀ relative to the untreated LD₉₀, and is a more operationallyrelevant measure of efficacy than the PR, especially if the slopes ofthe dose-lethality curves are significantly different. Militaryrequirements documents prescribe 80-90% survival for acceptance of newmedical countermeasures against nerve agent intoxication.

Results:

FIG. 1 graphs the probit dose-lethality curves for VX in composition andRSDL-decontaminated animals, and Table 1 summarizes the results based onLD₅₀s. A total of 15 and 26 animals were needed to generate thedose-lethality curves for RSDL and the composition, respectively, usingthe stopping criteria described in the methodology. The 24 hr dermalLD₅₀ of VX was 5959 μg/kg in animals decontaminated with the compositionand 3380 μg/kg in animals decontaminated with RSDL. The composition was1.8-fold (p<0.05) more effective than RSDL. The slope of the compositiondose-lethality curve was significantly (p<0.05) different from the slopeof the RSDL dose-lethality curve. The estimated PR (treated tountreated) was 42.6 for the composition and 24.1 for RSDL.

TABLE 1 Twenty-four hour VX _(LD50) Estimates in Guinea PigsDecontaminated with the Composition or RSDL 2 Min After Dermal ExposureSlope of the 24 hr VX LD₅₀ Estimated Number of Dose-Lethality μg/kg,p.c. Protective DC Product Animals Curve (95% CI) Ratio¹ Composition 26 6.4 5959 42.6 (4858-7309) RSDL 15 12.7 3380 24.1 (2921-3910) EfficacyRatio Composition/RSDL= 1.8 p <0.05 50% Survival Estimated using a24-hour dermal VX LD₅₀ of 140 μg/kg in fur-clipped unanesthetized guineapigs (Clarkson, personal communication)

Table 2 summarizes the results based on LD₁₀s. The 24 hr dermal LD₁₀ ofVX was 3755 μg/kg in animals decontaminated with the composition and2681 μg/kg in animals decontaminated with RSDL. The composition was1.4-fold more effective than RSDL; this difference was not significant.Also, presented in Table 2 is the ratio of the VX LD₁₀ in animalsreceiving DC to the VX LD₉₀ in animals that were not treated with a DCproduct. The LD₁₀/LD₉₀ ratio for the composition was 20 and the ratiofor RSDL was 14.

TABLE 2 Twenty-Four Hour VX LD₁₀ Estimates in Guinea Pigs Decontaminatedwith the Composition or RSDL 2 Min After Dermal Exposure 24 hr VX LD₁₀Number of μg/kg, p.c. DC Product Animals (95% CI) LD₁₀/LD₉₀ ₁ Compositon26 3755 20 (2390-5500) RSDL 15 2681 14 (2096-3429) Efficacy RatioComposition/RSDL = 1.4 90% Survival VX LD₉₀ of 188 μg/kg was used forthe denominator. This value was estimated from the dose-lethality alivegenerated in fur-clipped, unanesthetized guinea pigs (Clarkson, personalcommunication)

A comparison of the LD₅₀ estimates showed that the composition wassignificantly more effective than RSDL. In addition, the slope of thecomposition dose-lethality curve was more shallow than the slope of theRSDL curve. It is not unusual for the slope of the dose-lethality curveto become more shallow as the effectiveness of medical countermeasuresagainst organophosphate intoxication increases. However, Braue et al.observed no difference in the slopes of the dose-lethality curves forRSDL, 1% soapy water, and 0.5% bleach, even though RSDL was greater than3-fold more effective than the other two DC products; all three slopeswere similar to the slope for the composition in the study.

When the slopes are different, comparison of LD₅₀s may not be asvaluable, because the lower doses of agent in the curve with theshallower slope may still show lethality. Since the slope of thecomposition dose-lethality curve was shallower than the slope of theRSDL curve, the ratio of the LD₁₀ doses of VX were comprised. This mightreveal whether the shallower slope of the composition dose-lethalitycurve resulted in higher lethality at lower doses of VX compared toRSDL. The LD₁₀ was selected because military requirements documentsprescribe 80-90% survival rates as criteria for accepting new medicalcountermeasures for use by warfighters. The composition was still moreeffective than RSDL, but the ratio of the LD₁₀s was not significantlydifferent. This was probably due to the wider confidence intervalsaround the LD₁₀ than the LD₅₀ estimate. The ratio of the LD₁₀ in theanimals receiving DC to LD₉₀ in animals not receiving DC providesanother way of comparing efficacy which is independent of the slope.This ratio value represents the number of LD₉₀s of exposure that can betolerated without sustaining more than 10% lethality. This value was 20for the composition and 14 for RSDL.

Example 2

This example involved an In vitro evaluation of the composition of theinvention by nuclear magnetic resonance (NMR) evaluation. Thecomposition contained a concentrated solution of 10.7 wt % peraceticacid, 19.8 wt % hydrogen peroxide and 16.8 wt % acetic acid.

Summary: A decontamination (DC) solution containing the composition wasexamined for its ability to breakdown intact chemical warfare agents(CWAs) in vitro using nuclear magnetic resonance spectroscopy (NMR).Agents examined were HD, GD, VX, VR and A-232. For all agents except HD,the assessment was done with one-dimensional heteronuclear spin quantumcorrelation (HSQC) techniques. This approach has been successfullyutilized for similar studies examining the breakdown kinetics in enzymesystems using comparable agent levels. For HD, direct one-dimensionalproton experiments were used.

Composition Sample Preparation: Dilutions of the composition were madewith 99.5% D₂O from Sigma-Aldrich. Final concentrations of 2 wt %%, 4 wt% and 6 wt % of peracetic acid were achieved after mixing 100 μL of CWAwith 500 μL of the composition.

Agent Preparations: CWAs were provided in deuterated solvents by theMRICD Chemical Exclusion Area where the concentrations of each of theCWAs were determined independently. For all experiments, 100 μL of CWAwas mixed with 500 μL of the composition. For kinetic determinations,the peak area at time zero was set equal to the final amount of CWAafter dilution with the composition. Time zero amounts were 912.8,175.7, 95.3, 88.0 and 77.7 for HD, GD, VX, VR and A-232, respectively.

Instrumentation: NMR data on VX, VR, GD and A-232 were collected on a3-channel Bruker Avance III Ultrashield 500 MHz NMR spectrometer (BrukerBiospin, Billerica, Mass.) equipped with a Z-gradient 5 mm BBO probehead at 25° C. Topspin (Bruker 3.2pl6) was used for data acquisition andprocessing. Dynamics Center v2.2 was used for kinetic analysis, andresults were exported to PDF and Excel formats, which later wereanalyzed using GraphPad's Prism5 for Windows.

NMR data on HD were collected on an Agilent (Agilent Systems, SantaClara, Calif.) 4-channel DD2 Actively Shielded 600 MHz NMR instrumentequipped with a 5 mm PFG Penta probe at 25° C. VNMRJ 3.2 was used fordata collection and processing, and the results were exported to Excelfor kinetic analysis.

Experimental Design: For all experiments 100 μL of CWA was added to 500μL of the composition or D2O as per experimental protocol. Protonone-dimensional experiments were carried out with or withoutpre-saturation of water signal. The offset carrier frequency wasadjusted on water resonance, and minimum power level was used tosaturate the water signal without distortion/perturbation of theneighboring resonances. All samples were temperature equilibrated, andthe probe was tuned to the respective frequencies followed by gradientshimming. The time required for gradient shimming (dead time) was notedand taken into account for all kinetic calculations. Ninety degree pulsewidth, X pwx, and decoupling calibrations were done on both the protonand phosphorus channels and saved in the probe files, respectively.Proton one-dimensional pulse sequence (s2pul) was used in HD, andmodified gradient ¹H-X 1D-HSQC (FIG. 2) was used for all other CWAs. (Arepresentation of the gHSQC1D experiment is presented in FIGS. 3a and 3bshowing expansion of the region of interest of methyl resonances —P—CH₃at 1.96 and 1.20 ppm, respectively.) Optimization of the proton signalat 1.96 ppm in ¹H-X Heteronuclear Single Quantum Coherence (HSQC) wasdone by observing ^(n)J_(HX) coupling constant (Table 3) and changingthe J parameter in the pulse sequence to the observed value. The ratioof G1/G2 gradient strength was also optimized to give an optimum signal,and this is based on the gyromagnetic ratio of proton and phosphorus.

TABLE 3 Coupling Constants for Various Chemical Warfare Agents ^(n)JHXCoupling Constant (Hz) ²JHX VX 17.5 ²JHX GD 18.5 2J HX VR 17.5

Proton spectra were acquired on all CWAs to check the purity of thecompound and to determine whether any degraded products were present.Background spectra were acquired on all composition samples before 100μL of respective agent was added to the NMR tube. All gHSQC1D spectrawere acquired with the same parameters (nt/ns=16; relaxation delay(d1)=2 sec; dummy scans (ss/ds)=0; acquisition time (at)=1.7s; number ofdata points (np)=32K, Fourier number (fn)=64K) and processed usingBruker Biospin and/or VNMRJ with zero filling to 64K data size andapplying exponential weighting function line broadening (LB) of 1.0 Hz.

Peak integrals were manually defined based on methyl peaks from startingmaterial and methyl peaks from intermediate compound. These methyl peakswere identified by using an edited version of (gradient heteronuclearsingle quantum coherence with adiabatic pulses) gHSQCAD two-dimensionalexperiment and the peaks' chemical shifts. Total acquisition time forall kinetic experiments was adjusted to 1 hour. Triplicate data setswere collected for each of the four CWA and blank runs. Blank runs weredone in 99.8% D2O without the composition and monitored for a total of 1hr. Kinetic data are summarized in Tables 4 and 5.

TABLE 4 Half-Lives of CWAs Decontaminated with the composition Solutions2% 4% 6% Blank Composition Composition Composition RSDL T½ (min) (95%Confidence Interval) HD 8.6 51.6 27.9 19.7 NA (8.4-8.8) (51.4-51.7)(27.7-28.1) (19.55-19.79) GD N/A 1036 797 915 NA (907-1210) (709-910)(680-1398) VX N/A 39.7 19.2 13.7 4.6 (38.5-41.0) (18.9-19.5)(13.43-13.89) (4.4-4.9) VR N/A 30.0 14.9 12.5 NA (29.3-30.7) (14.6-15.1)(12.3-12.8) A-232 N/A 163 95.3 54.3 NA (135-206) (85-108) (50.6-58.6)

TABLE 5 Percentage of CWA Remaining after 60 Min Decontamination withthe Composition Solution 2% 4% 6% Blank Composition CompositionComposition HD 0 45 24 13 GD 97 93 90 95 VX 100 34 11 8 VR 100 27 7 5A-232 100 90 61 48 *Decomposition of HD in the blank was due tohydrolysis, while oxidative breakdown was the major route observed inthe composition.

To determine the rate of hydrolysis of the CWAs in the reaction mixture,the gHSQC method was used, and the peak corresponding to the startingmaterial was manually integrated. This integration area was then appliedto all spectra to obtain the change in concentration over time. In thecase of HD, one-dimensional proton spectra were used. The peakcorresponding to the methylene (—CH2-) peak of the starting material wasintegrated manually and then was applied to all spectra collected.

Results: In blank composition solution (water), HD rapidly hydrolyzedwith a t1/2 of 8.6 minutes (Table 4 and FIG. 4). In contrast, GD, VX, VRand A-232 were found to be stable, demonstrating that 97-100% of thestarting concentration remained at the end of the experiment (Table 5).In the composition, HD breakdown was slower at all concentrationsrelative to the blank. Half-lives were approximately 52, 28 and 20minutes for 2, 4 and 6%, respectively (Table 4). The differentialbreakdown kinetics observed between the blank and the compositionsolutions can be explained by the products formed. Breakdown productsidentified in the blank indicate hydrolysis as the primary route,whereas in the composition solutions, oxidative processes were involved.The composition demonstrated no decontamination efficacy for GD (Table4). The amount remaining at the end of the experiment was 90% or greaterat all the composition concentrations (Table 5). Aconcentration-dependent breakdown was observed for VX (FIG. 5) and VR inthe composition. Half-lives were 39, 19 and 13 minutes for 2, 4 and 6%,respectively, for VX and 30, 15 and 12 minutes for 2, 4 and 6%,respectively, for VR (Table 4). Some breakdown of A-232 was observed inthe composition compared to blank, but the rates were much less thanthose observed for either VX or VR in the composition (Table 4). Thekinetic data are summarized in Tables 4 and 5.

Conclusion: At the time of writing, measurements by NMR of the rate atwhich RSDL breaks down CWAs had only been done for RSDL vs VX. It wasdetermined that RSDL breaks down VX with a half-life of 4.6 minutes.Although the composition was slower than RSDL at breaking down VX, itwas fast enough to support testing the composition in vivo.

Example 3

This example involved an initial skin DC efficacy evaluation of thecomposition and its comparison to RSDL following skin application of VXin guinea pigs.

Animals: The same type of animals were secured and prepared as describedin Example 2.

Materials: An aliquot of neat VX was obtained from the USAMRICD ChemicalExclusion Area each exposure day. RSDL was purchased in sealed packagesfrom First Line Technology, Chantilly, Va. The composition, containing aconcentrated solution of 10.7 wt % peracetic acid, 19.8 wt % hydrogenperoxide and 16.8 wt % acetic acid, and an aliquot was diluted to a 2%peracid concentration in deionized water each test day.

Agent exposure: A vial of neat VX was obtained from the USAMRICDChemical Exclusion Area each exposure day and placed in a fume hood atroom temperature. VX was applied with a Hamilton 0.5 ul, 1 ul or 5 ulsyringe equipped with a blunt tip needle and digital dispenser to theoutlined exposure area.

DC Procedures: Decontamination procedures were conducted in the samemanner described in Example 2.

Experimental Design: Experimental data were generated as described inExample 2.

Statistical Analysis: A final probit analysis was conducted on allstages from the 24 hr responses for each DC material. The slopes andLD50s as well as the LD1, LD10, LD16, LD30, LD70, LD84, LD90, and LD99were calculated with their respective Fieller's and Delta 95% confidenceintervals (CI). LD50 estimates for RSDL and the composition werecompared using another specialized SAS program (Battelle, Columbus,Ohio), which determined whether the ratio of the LD50s was statisticallydifferent at p<0.05. A significant (p<0.05) difference was achieved whenthe delta 95% CI of the LD50 ratio did not include the value of 1.⁵ Aprotective ratio (PR) defined as the LD50 of VX in animals treated withthe DC product divided by the LD50 of VX in untreated animals wasestimated, using a historic value of 140 μg/kg⁸ in fur-clippedun-anesthetized guinea pigs for the denominator in the ratio.

Results: FIG. 6 graphs the probit predicted dose-lethality curves for VXin the composition- and RSDL-decontaminated animals and tabulates theprobit estimates at 24 hr for the LD₅₀, 95% CI and slope. A total of 15and 26 animals were needed to generate the dose lethality curves forRSDL and the composition, respectively, using the stopping criteriadescribed above. The 24 hr dermal LD₅₀ of VX was 5959 μg/kg in thecomposition-decontaminated animals and 3380 μg/kg in RSDL-decontaminatedanimals. The composition was 1.8-fold (p<0.05) more effective than RSDL.The estimated PR (treated/untreated) was 42.6 for the composition and24.1 for RSDL. Since the gauze used with the composition was moreabrasive than the sponge used with RSDL, the possibility that the higherlevel of protection provided by the composition may have been due toincreased physical removal.

Conclusion: The results of this initial assessment of the efficacy ofthe composition suggest that it is more effective than RSDL as a skin DCproduct for the nerve agent VX. However, the RSDL PR in this study isconsiderably lower than PRs reported by either Braue et al. or Clarksonet al.8 These investigators reported RSDL PRs of 66 (N=37) and 52(N=53), respectively, using the same DC timing and procedures. We haveno explanation for the difference in their results from ours except forthe smaller sample size used in the current experiment and the inherentvariability in responses of animals.

Example 4

This example analyzed delayed skin DC efficacy of the composition andRSDL following topical A-232 application in guinea pigs.

Animals: Animal information is the same as in previous studies.

Materials: An aliquot of neat A-232 was obtained from the USAMRICDChemical Exclusion Area each exposure day. RSDL, purchased in sealedpackages from First Line Technology, Chantilly, Va. The containing aconcentrated solution of 10.7 wt % peracetic acid, 19.8 wt % hydrogenperoxide and 16.8 wt % acetic acid was prepared. Aliquots were dilutedto a 2% peracid concentration in deionized water according to themanufacturer's formula each test day.

Agent Exposure: Exposure methods for A-232 were the same as thosedescribed for VX in previous studies.

DC Procedure: RSDL and composition applicators and DC procedures werethe same as described in previous studies. Water was applied with gauzeapplicators saturated with 10 ml using the same procedures utilized forRSDL and the composition. A fresh applicator of each DC product was usedon each animal.

Experimental Design: This study consisted of two experiments, each with30 animals. Animals in each experiment were randomly allocated intothree DC treatment groups of 10; the groups were RSDL, 2% composition,and water. In Experiment 1, animals were exposed topically to 5×LD₅₀s ofneat A-232, and skin DC was performed at 3 hr after agent application orat the onset of signs, whichever occurred first. In Experiment 2,animals were exposed topically to 10×LD₅₀ of neat A-232, and skin DC wasperformed at 1 hr after agent application or at the onset of signs,whichever occurred first. The clinical condition of each animal wasevaluated at 2 hr and 4 hr after DC, early the next morning (0700-0800)and at 24 hr after exposure. Clinical assessment scores (CAS) were givento the animals at each observation time according to the followingscale: 0=normal appearance and behavior; 1=mildly intoxicated(characterized by slight lethargy and/or minor signs, but animal isupright and ambulates by itself); 2=moderately intoxicated (noticeablelethargy and signs, but animal is upright and will ambulate if prodded);3=severely intoxicated (the animal is prone, not ambulatory, withprominent signs but is conscious and can lift head); and 4=very severelyintoxicated (animal is prostrate, unresponsive with or withoutpronounced signs).

Results: Experiment 1: Six of the 30 animals developed signs of nerveagent intoxication prior to the three-hour DC time. All but one of thesesix animals were decontaminated at the onset of signs with theirassigned DC product, and all subsequently died prior to 24 hr. The oneanimal that was not decontaminated developed signs within 10 min afterexposure and was dead by 13 min after exposure. The remaining 23 animalsreached the 3 hr DC time without exhibiting any visible signs of nerveagent intoxication. Each was decontaminated, and the survival resultsare summarized in Table 6. The survival rates for the RSDL, composition(2%), and water DC groups were 9/9 (100%), 7/8 (88%) and 4/6 (67%),respectively. One animal in the composition DC group was removed fromthe study due to a technical error.

TABLE 6 Summary of Survival from Experiment 1 Survival Rate DC Product 2hr 4 hr Next AM 24 hr RSDL 9/9 9/9 9/9 9/9 Composition (2%) 8/8 8/8 7/87/8 Water 6/6 6/6 5/6 5/6

Table 7 shows the clinical assessment scores at 2 and 4 hr after DC, thenext morning, and at 24 hr after exposure in the subset of 23 animalsthat were sign-free when DC was performed 3 hr after dermal exposure.Only one animal exhibited signs at 2 hr; this was an RSDL animal. Theremaining animals that developed signs did so between 4 hr after DC andthe next morning, at which time one composition animal was found dead,and two of the animals in the water group were severely affected. At 24hr, 4/9 RSDL-decontaminated animals were normal and the remaininganimals were scored as mild to moderately affected; 4/7 survivingcomposition animals were normal looking, and the remaining animals weremild to moderately affected. In the water DC group at 24 hr, 1/4surviving animals appeared normal, two animals died, and three were mildto moderately affected.

TABLE 7 Clinical Assessment Scores in Individual Animals at 2 and 4 Hrafter DC , the Next Morning and at 24 Hr after Exposure to 5xLD₅₀ A-232(Experiment 1) Clinical Assessment Scores DC Product Animal # 2 hr 4 hrNext AM 24 hr RSDL 288 0 N/A 1 2 291 0 N/A 0 1 294 0 N/A 0 1 297 0 0 0 0299 0 0 0 0 301 0 0 0 1 308 0 0 0 0 311 2 2 1 2 313 0 0 0 0 Composition(2%) 287 0 N/A 0 1 292 0 N/A Dead Dead 293 0 N/A 1 2 296 0 N/A 0 0 305 00 1 1 309 0 0 0 0 314 0 0 0 0 315 0 0 0 0 Water 289 0 N/A 1 0 290 0 N/A1 2 304 0 0 3 Dead 307 0 1 3 Dead 310 0 0 1 1 312 0 1 1 2

Experiment 2: Three of the 30 animals developed signs prior to 1 hr. All3 were decontaminated with their assigned DC product at the onset ofsigns, and all died within 5 min after DC. The remaining 27 animalsreached the one-hour DC time without exhibiting any signs ofintoxication. Each was decontaminated, and the survival results aresummarized in Table 8. The 24 hr survival rates were 9/9 (100%), 9/10(90%) and 6/8 (75%) in the RSDL, the composition and water DC groups,respectively.

TABLE 8 Summary of Survival from Experiment 2 Survival Rate DC Product 2hr 4 hr Next AM 24 hr RSDL 9/9 9/9 9/9 9/9 Composition (2%) 10/10 10/10 9/10  9/10 Water 8/8 8/8 6/8 6/8

Table 9 shows the clinical assessment scores at 2 and 4 hr after DC, thenext morning, and at 24 hr after exposure in the subset of animals thatwere sign-free when DC was performed 1 hr after exposure. Over thecourse of the post-DC period, 5 of 9 RSDL animals, 6 of 10 compositionanimals and 6 of 8 water animals displayed signs of nerve agentpoisoning of varying severity and time to onset. At 24 hr, none of theRSDL animals had died, one was very severely intoxicated, and 8 of 9were normal looking. In the composition group, one animal died, twoother animals had mild or moderate signs and 7 of 10 were normallooking. The water-decontaminated animals at 24 hr were clearly sickerthan the animals in the composition and RSDL groups. Only 2 of 8 animalswere normal looking, two had died, two were severely or very severelyaffected and two had mild to moderate signs.

TABLE 9 Clinical Assessment Scores in Individual Animals at 2 and 4 Hrafter DC, the Next Morning and at 24 Hr after Exposure to 10xLD₅₀(Experiment 2) Clinical Assessment Scores (CAS) DC Product Animal # 2 hr4 hr Next AM 24 hr RSDL 318 1 0 0 0 321 0 0 0 0 323 1 0 1 0 327 0 0 0 0330 0 0 0 0 333* 1 0 0 0 336* 3 3 4 4 340* 0 0 0 0 344* 2 2 0 0Composition 319 0 0 0 0 (2%) 324 0 1 1 1 325 0 0 2 2 328 0 0 0 0 331 0 0Dead Dead 332* 0 0 0 0 334* 2 0 0 0 335* 1 0 0 0 343* 0 0 0 0 345* 1 0 00 Water 317 0 0 0 0 320 1 1 Dead Dead 322 0 0 2 3 326 0 1 2 1 329 0 0 34 337* 1 2 Dead Dead 339* 0 0 0 0 346* 1 2 2 2

Conclusion: RSDL and the composition were both effective in preventinglethality when used 3 hr after dermal application of 5×LD₅₀ of thisagent in un-anesthetized, fur-clipped animals. Each group had highsurvival rates at 24 hr. Mild to moderate toxic signs were present in5/9 RSDL animals and in 4/8 of the composition animals.

The water DC group was included in the study as a control to evaluatethe role of physical removal. Survival rate (83%) at 24 hr in the watergroup was not different from the RSDL and composition animals; however,more of the water-decontaminated animals died, showed signs ofintoxication, and were more severely affected the day after exposure.

Example 5

This example analyzed skin decontamination efficacy of the compositionand RSDL in guinea pigs following topical VX application, using methodsthat mimic mass casualty DC procedures

Animals: Animal information is the same as in previous studies.

Materials: An aliquot of neat VX was obtained from the USAMRICD ChemicalExclusion Area each exposure day. RSDL sponge pads were purchased insealed packages from First Line Technology, Chantilly, Va. Bulk RSDL waspurchased from Emergent Biosolutions, Rockville, Md. The composition wasprepared as a concentrated solution of 9.5 wt % peracetic acid, 17.4%hydrogen peroxide and 17.8 wt % acetic acid, and aliquots were dilutedto a 2% peracid concentration in deionized water each test day.

Agent Exposure: Agent exposure method was the same as in previous VXstudies.

Decontamination (DC) Procedure: The exposure site on each animal wasdecontaminated two minutes after VX application with tap water, 1% Dawn™dish detergent, RSDL, or 2% composition. A three-step DC proceduredesigned to mimic procedures employed in a mass casualty chemical agentincident was utilized. In step 1, the DC material was applied by swipingan applicator containing the DC material 10 times quickly in shortstrokes across the exposure site in a head-to-tail direction; the DCmaterial remained on the skin for 2 min. In step 2, the DC material wasremoved by wiping the exposure site 5 times quickly in short strokes ina head-to-tail direction with an applicator wetted with 10 ml of water.In step 3 the exposure area was dried with 5 swipes with a dry gauzeapplicator. Fresh applicators were used for each procedure on eachanimal. Water, Dawn™, and the composition were applied with fresh gauzeapplicators for each animal saturated with 10 ml of DC solution, asdescribed in previous experiments. RSDL was applied using either thegauze applicator (RSDL-G) saturated with 10 ml of RSDL or the commercialsponge pad applicator (RSDL-S), as described in previous studies.

Experimental Design: VX dose-lethality curves (DLCs) were generated forall DC materials using a modified stage-wise adaptive dose design.1 Eachstage consisted of a number of agent doses and animals per dose toestablish the range of lethality from 0-100% and to define the responserelationship in the projected middle (30-70%) of the DLC. A specializedprobit analysis program (Battelle, Columbus, Ohio) and SAS NLIN wereused on the cumulative results (survival/lethality) after each stage toestimate the LD₅₀ and 95% confidence intervals (CI) and to assessstopping criteria to limit animal use. The stage process continued withadditional challenge doses and animals per agent dose until the ratio ofthe upper 95% CI minus the lower 95% CI (delta or Fieller's limits)divided by two times the MLD estimate from the latest stage was lessthan 0.4 (stopping criteria) or up to 40 animals were used. Afterexposure and DC, each animal was monitored continuously for up to 2 hruntil the onset of toxic signs appeared, then at 4 hr after DC and againat 24 hr after exposure.

Statistics: A final probit analysis was conducted on all stages from the24 hr responses for each DC material. The LD50s, LD1, LD10, LD16, LD30,LD70, LD84, LD90, and LD99 with their respective 95% CI were calculatedby both Fieller's and delta methods, and the slopes were determined.3,4A protective ratio (PR), defined as LD50 of VX in animals treated witheach DC material divided by the historic LD50 of VX in untreated animalsof 140 μg/kg8 in fur-clipped un-anesthetized guinea pigs, wascalculated. This estimate was used to compare the effectiveness of theDC treatments using a specialized probit program (Battelle, Columbus,Ohio) and SAS.

Results: FIG. 7 graphs the 24 hr probit-predicted VX dose-lethalitycurves for skin decontamination at 2 min after agent application withwater, 1% Dawn′, RSDL-G, RSDL-S and 2% composition using a 3-step DCprocedure as well as the probit estimates at 24 hr for the LD50, 95% CIand slope for each DC material. The composition was significantly moreeffective than any of the other DC materials, with an estimated PR of108. The composition was more than 4 times more effective than tap wateror soapy water, and more than 2 times more effective than RSDL appliedwith the commercial sponge pad (RSDL-S) or with the gauze (RSDL-G)applicator. There was a trend suggesting that the RSDL-S was moreeffective than RSDL-G.

Conclusion: The composition was at least 2-fold more effective than RSDLand 4- to 4.5-fold more effective than tap water or soapy water using a3-step skin DC methodology that mimics mass DC procedures againsttopically applied VX in guinea pigs. Since the composition was appliedand left on the skin for only 2 min prior to rinsing and drying of theskin, the results suggest that the composition neutralized VX on skin.

Example 6

This example analyzed Skin DC efficacy of the composition and RSDLfollowing topical VX application in swine.

Animals: Gottingen mini-pigs were acclimated to being in a transfersling and in a cage in a fume hood. Animals were trained to respond toGatorade, which was used to entice the animal to approach the edge ofthe cage with its head down so that agent could be applied to the scalpand decontamination solution could be rubbed across the exposure site.On the day of exposure, the animals were weighed, the hair on the scalpwas clipped and a 1-inch diameter circle was drawn on the scalp.

Experimental Design: While the animal was being given Gatorade, 2×LD₅₀(490 μg/kg) of VX was applied using a Hamilton digital syringe to thecenter of the circle. At either 5 min or 1 hour after VX application,the site was decontaminated with either RSDL or the composition, bywiping the site 3 times with the DC product. Care was taken to ensurethat the DC product did not enter the eye by using a piece of dry gauze.The DC product was allowed to stay on the skin for 15 minutes, thenremoved by wiping the site 3 times with saline-soaked gauze. Animalswere observed and kept in the hood for 24 hours, with access to food andwater. All surviving animals were euthanized at 24 hours. The exposurearea of the scalp was removed and placed in bleach for decontamination.

Conclusion: In this mini-pig model, RSDL and the composition werecomparable in decontamination effectiveness.

Many modifications and other examples of the disclosure set forth hereinwill come to mind to those skilled in the art to which this disclosurepertains, having the benefit of the teachings presented in the foregoingdescriptions and the associated drawings. Therefore, it is to beunderstood that the disclosure is not to be limited to the specificexamples disclosed and that modifications and other embodiments areintended to be included within the scope of the appended claims.

Moreover, although the foregoing descriptions and the associatedembodiments describe aspects of the disclosure in the context of certainexample combinations of structural elements and/or functions, it shouldbe appreciated that different combinations of elements and/or functionsmay be provided by alternative embodiments without departing from thescope of the appended claims. In this regard, for example, differentcombinations of elements and/or functions than those explicitlydescribed above are also contemplated as may be set forth in some of theappended claims. Although specific terms are employed herein, they areused in a generic and descriptive sense only and not for purposes oflimitation.

What is claimed is:
 1. A chemical warfare agent decontaminatingcomposition comprising: one or more peroxyacids in an amount rangingfrom about 0.02% to about 10% by weight; one or more hydroperoxides inan amount ranging from about 0.01% to about 25% by weight; and one ormore C₂₋₁₀ carboxylic acids in an amount ranging from about 0.01% toabout 30% by weight, wherein at least one of the carboxylic acids is thecorresponding carboxylic acid of the peroxyacid; and wherein thecomposition is formulated in the form of an aerosol or liquid spray or agel, sol, liquid, lotion or irrigation gel, optionally saturated on anabsorbent applicator comprising an absorbent sponge, natural orsynthetic, woven or non-woven fabric, cloth, paper towel or towelette.2. The decontaminating composition of claim 1, wherein the one or morehydroperoxides comprise hydrogen peroxide.
 3. The decontaminatingcomposition of claim 1, wherein the amount of the hydroperoxide rangesfrom about 0.1% to about 20% by weight.
 4. The decontaminatingcomposition of claim 1, wherein the amount of the carboxylic acid rangesfrom about 0.1% to about 22% by weight.
 5. The decontaminatingcomposition of claim 1, wherein the one or more carboxylic acidscomprise acetic acid.
 6. The decontaminating composition of claim 1,wherein the one or more peroxyacids comprise peracetic acid.
 7. Thedecontaminating composition of claim 1, wherein the peroxyacid consistsessentially of peracetic acid and acetic acid is the correspondingcarboxylic acid.
 8. The decontaminating composition of claim 1, whereinthe composition further comprises a stabilizer selected from the groupconsisting of etidronic acid, di sodium ethylenediamine-tetraaceticacid, ethylenediaminetetraacetic acid, tetrasodiumethylenediaminetetraacetate, citramalic acid, dipicolinic acid,dipicolinic acid N-oxide, or a combination thereof.
 9. Thedecontaminating composition of claim 1, further comprising a magnesiumsalt.
 10. The decontaminating composition of claim 9, wherein themagnesium salt is the magnesium salt of peracetic acid.
 11. Thedecontaminating composition of claim 10, wherein the magnesium salt ismagnesium acetate tetrahydrate.
 12. The decontaminating composition ofclaim 1, wherein the composition comprises: peracetic acid in an amountranging from less than 1% to about 6% by weight; hydrogen peroxide in anamount ranging from about 0.1% to about 20% by weight; and acetic acidin an amount ranging from about 0.1% to about 20% by weight.
 13. Thedecontaminating composition of claim 1, wherein the compositioncomprises: peracetic acid in an amount ranging from about 0.1% to about6% by weight; hydrogen peroxide in an amount ranging from about 0.1% toabout 12% by weight; and acetic acid in an amount ranging from about0.1% to about 12% by weight.
 14. The decontaminating composition ofclaim 1, further comprising a surfactant.
 15. The decontaminatingcomposition of claim 14, wherein the surfactant is sodium dodecylsulfate.
 16. A method of decontaminating the skin, or a wound thereon,of an animal exposed to a chemical warfare agent, comprising contactingthe exposed skin or wound with an amount of a decontaminatingcomposition that is sufficient to decontaminate the skin or wound,wherein the decontaminating composition comprises: one or moreperoxyacids in an amount ranging from about 0.02% to about 10% byweight; one or more hydroperoxides in an amount ranging from about 0.01%to about 25% by weight; and one or more C₂₋₁₀ carboxylic acids in anamount ranging from about 0.01% to about 30% by weight, wherein at leastone of the carboxylic acids is the corresponding carboxylic acid of theperoxyacid.
 17. The method of claim 16, wherein the chemical warfareagent is selected from the group consisting of a V-series nerve agent,G-series nerve agent, A-series nerve agent, mustard agent, or acombination thereof.
 18. The method of claim 16, wherein the compositionis formulated in the form of an aerosol or liquid spray or a gel, sol,liquid, lotion or irrigation gel, optionally saturated on an absorbentapplicator comprising an absorbent sponge, natural or synthetic, wovenor non-woven fabric, cloth, paper towel or towelette.
 19. The method ofclaim 18, wherein applicator is in the form of a mitt.
 20. The method ofclaim 16, wherein the decontamination composition is contacted with theskin or wound by at least one of: soaking the skin or wound in asolution of the composition, dispensing a pressurized solution of thecomposition onto the skin or wound, or applying a coating or layer ofthe composition on skin or wound.
 21. The method of claim 19, whereinthe coating or layer is applied with the absorbent applicator.
 22. Themethod of claim 16, wherein the composition further comprises asurfactant.
 23. The method of claim 16, wherein the animal is a human.24. A kit comprising the composition of claim 1, packaged with anabsorbent applicator for the composition or medical countermeasures forchemical weapons packaged for administering to an exposed subject. 25.The kit of claim 24, wherein the absorbent applicator comprises anabsorbent sponge, natural or synthetic, woven or non-woven fabric,cloth, paper towel or towelette.
 26. The kit of claim 24, wherein theapplicator is saturated with the composition prior to packaging.
 27. Thekit of claim 24, wherein the applicator is a mitt.
 28. The kit of claim24, further comprising a container for safe disposal of usedapplicators.
 29. The kit of claim 24, wherein the medical countermeasureis a nerve agent antidote packaged for administering to an exposedsubject.
 30. The kit of claim 24, wherein an applicator is saturated andcontained inside of a bottled filled with the composition.